Abstract
Present study aimed to evaluate the influence of carbon/nitrogen ratio (C/N) on mixotrophic growth of microalgae and role of nanomaterial in cell recovery and lipid improvement. In this study, three microalgae species were isolated, screened from local freshwater body for lipid assimilation. The microalgae were identified as Chlorococcum sp., Scenedesmus sp., and Euglena sp. Mixotrophic cultivation of each microalgae strain using various organic carbon sources was preferred in contrast with photoautotrophic mode. Sucrose represented as the preeminent source for enhancing the microalgae biomass of 3.5 g/L and lipid content of 58.35%, which was a significant improvement as compared to control. Later, response surface methodology–central composite design (RSM–CCD), tool was employed to optimize the C/N ratio and demonstrated the maximum biomass production of 5.02 g/L along with the increased lipid content of 60.34%. Ti nanoparticles (Ti nps) were added to the culture for lipid enhancement in the stationary phase and biomass removal was performed by nanoparticle (np)-mediated flocculation technique. Optimized concentration of 15 ppm Ti nps determined the cell harvesting efficacy of 82.46% during 45 min of sedimentation time and 1.23-fold lipid enhancement was reported. Extracted lipid was converted to fatty acid methyl esters (FAME) by the process of transesterification and analyzed by gas chromatography–mass spectrometry (GC–MS). Characterization of FAME revealed the presence of 56.31% of saturated fatty acid (SFA) and 29.06% unsaturated fatty acids (UFA) that could be processed towards sustainable biodiesel production. Hence, our results suggested that integration of mixotrophic cultivation and Ti nps emerged as a new cost-effective approach for biomass and lipid enhancement in microalgae Chlorococcum sp.
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References
Anahas AM, Muralitharan G (2019) Central composite design (CCD) optimization of phytohormones supplementation for enhanced cyanobacterial biodiesel production. Renew Energ 130:749–761
Ansari FA, Shriwastav A, Gupta SK, Rawat I, Bux F (2017) Exploration of micro-algae biorefinery by optimizing sequential extraction of major metabolites from Scenedesmus obliquus. Ind Eng Chem Res 56(12):3407–3412
Arora N, Patel A, Sharma M, Mehtani J, PruthiPA PV, Poluri KM (2017) Insights into the enhanced lipid production characteristics of freshwater microalgae under high salinity conditions. Ind Eng Chem Res 56(25):7413–7742
Bartley ML, Boeing WJ, Daniel D, Dungan BN, Schaub T (2015) Optimization of environmental parameters for Nannochloropsis salina growth and lipid content using the response surface method and invading organisms. J Appl Phycol 28:15–24
Bligh EG, Dyer WJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem Phys 37:911–991
Chen J, Jiang X, Wei D (2020) Effects of urea on cell growth and physiological response in pigment biosynthesis in mixotrophic Chromochloris zofingiensis. J Appl Phycol. https://doi.org/10.1007/s10811-020-02114-3
Chia SR, Chew KW, Show PL, Yap YJ, Ong HC, Ling TC, Chang JS (2018) Analysis of economic and environmental aspects of microalgae biorefinery for biofuels production: a review. Biotechnol J 13(6):e1700618
Feng P, Xu Z, Qin L, Alam MA, Wang Z, Zhu S (2020) Effects of different nitrogen sources and light paths of flat plate photobioreactors on the growth and lipid accumulation of Chlorella sp GN1 outdoors. Bioresour Technol 301:122762
Fritsch FE (ed) (1948) The structure and reproduction of the algae. Cambridge, The University Press, pp 147–153
Gallagher BJ (2011) The economics of producing biodiesel from algae. Renew Energy 36:158–162
Garcia PF, Kubbutat P, Brammen M, Schwaminger S, Berensmeier S (2018) Bare iron oxide nanoparticles for magnetic harvesting of microalgae: from interaction behavior to process realization. Nanomaterials 8:292
He M, Yan Y, Pei F, Wu M, Gebreluel T, Zou S, Wang C (2017) Improvement on lipid production by Scenedesmus obliquus triggered by low dose exposure to nanoparticles. Sci Rep 7:15526
Hossain N, Mahila TKI (2019) Progress in physicochemical parameters of microalgae cultivation for biodiesel production. Crit Rev Biotechnol 39(6):835–859
Hossain N, Zaini J, Mahila TKI (2018) Experimental investigation of energy properties for Stigonematales sp. microalgae as potential biofuel feedstock. Int J Sustain Eng 12(2):123–130
Hossain N, Mahlia TMI, Saidur R (2019) Latest development in microalgae biofuel production with nano-additives. Biotechnol Biofuels 12:125
Hsu JP, Liu BT (1998) Critical coagulation concentration of a colloidal suspension at high particle concentrations. J Phy Chem B 102:334–337
Hu X, Liu B, Deng Y, Bao X, Yang A, Zhou J (2019) A novel two-stage culture strategy used to cultivate Chlorella vulgaris for increasing the lipid productivity. Sep Purif Technol 211:816–822
Hua L, Guo L, Thakkar M, Wei D, Agbakpe M, Kuang L, Magpile M, Chaplin BP et al (2016) Effects of anodic oxidation of a substoichiometric titanium dioxide reactive electrochemical membrane on algal cell destabilization and lipid extraction. Bioresour Technol 203:112–117
Huang WC, Kim JD (2016) Nickel oxide nanoparticle-based method for simultaneous harvesting and disruption of microalgal cells. Bioresour Techol 218:1290–1293
Jeyakumar K, Asha D, Varalakshmi P, Kathiresan S (2020) Nitrogen repletion favors cellular metabolism and improves eicosapentaenoic acid production in the marine microalga Isochrysis sp. CASA CC 101. Algal Res 47:101877
Jiang SY, Chi YH, Wang GZ, Zhou JX, Cheng YS, Zhang BL, Ma A, Vanitha J, Ramachandran S (2015) Sucrose metabolism gene families and their biological functions. Sci Rep 5:17583–17607
Khanra A, Vasistha S, Rai MP (2017) Glycerol on lipid enhancement and FAME characterization in algae for raw material of biodiesel. Int J Ren Energ Res 7(4):1970–1978
KhanraA SS, Shakeel A, Suhag D, Mistry S, Rai MP, Chakrabarti S, Mukherjee M (2018) Sustainable growth and lipid production from Chlorella pyrenoidosausing N-doped carbon nanosheets: unravelling the role of graphitic nitrogen. ACS Sust Chem Eng 6(1):774–780
Koley S, Prasad S, Bagchi SK, Mallick N (2017) Development of a harvesting technique for large scale microalgal harvesting for biodiesel production. RSC Adv 7:7227
Lee YC, Lee HU, Lee K, Kim B, Lee SY, Choi MH, Farooq W, Choi JS, Park JY, Lee J, Oh YK, Huh YS (2014) Aminoclay-conjugated TiO2 synthesis for simultaneous harvesting and wet-disruption of oleaginous Chlorella sp. Chem Eng J 245:143–149
Lin Q, Lin J (2011) Effects of nitrogen source and concentration on biomass and oil production of a Scenedesmus rubescens like microalga. Bioresour Technol 102:1615–1621
Lin TS, Wu JY (2015) Effect of carbon sources on growth and lipid accumulation of newly isolated microalgae cultured under mixotrophic condition. Bioresour Technol 184:100–107
Mondal M, Goswami S, Ghosh A, Oinam G, Tiwari ON, Das P, Gayen K, Mandal MK, Halder GN (2017) Production of biodiesel from microalgae through biological carbon capture: a review. 3 Biotech 7:99
Nayak M, Suh WI, Chang YK, Lee B (2019) Exploration of two-stage cultivation strategies using nitrogen starvation to maximize the lipid productivity in Chlorella sp. HS2. Bioresour Technol 276:110–118
Nguyen MK, Moon JY, Bui VKH, Oh YK, Lee YC (2019) Recent advanced applications of nanomaterials in microalgae biorefinery. Algal Res 41:101522
Rai MP, Gupta S (2017) Effect of media composition and light supply on biomass, lipid content and FAME profile for quality biodiesel production from Scenedesmus abundans. Energ Converg Manage 141:85–92
Rai MP, Nigam S, Sharma R (2013) Response of growth and fatty acid compositions of Chlorella pyrenidosa under mixotrophic cultivation with acetate and glycerol for bioenergy application. Biomass Bioenerg 58:251–257
Rai MP, Khanra A, Rai S, Srivastava M, Prakash R (2018) Pivotal role of levoglucosenone and hexadecanoic acid from microalgae Chlorococcum sp. for corrosion resistanceon mild steel: electrochemical, microstructural and theoretical analysis. J Mol Liq 266:279–290
Sehgal A, Goswami K, Pal M, Chikkaputtaiah C, Chetia P, Boruah HPD (2019) Morpho-taxonomic, genetic, and biochemical characterization of freshwater microalgae as potential biodiesel feedstock. 3 Biotech 9(4):137
Sharma AK, Sahoo PK, Singhal S, Patel A (2016) Impact of various media and organic carbon sources on biofuel production potential from Chlorella spp. 3 Biotech 6:116
Sun Y, Liu J, XieT XX, Liu W, Liang B, Zhang Y (2014) Enhanced lipid accumulation by Chlorella vulgarisin a two-stage fed-batch culture with glycerol. Energ Fuels 28:3172–3177
Tan KWM, Lin H, Shen H, Lee YK (2016) Nitrogen-induced metabolic changes and molecular determinants of carbon allocation in Dunaliella tertiolecta. Sci Rep 6:37235
Wang YZ, Hallenbeck PC, Leite GB, Paranjape K, Huo DQ (2016) Growth and lipid accumulation of indigenous algal strains under photoautotrophic and mixotrophic modes at low temperature. Algal Res 16:195–200
Yu Q, Wang H, Li X, Yin Y, Qin S, Ge B (2020) Enhanced biomass and CO2 sequestration of Chlorella vulgaris using a new mixotrophic cultivation method. Process Biochem 90:168–176
Zhu L, Li S, Hu T, Nugroho YK, Yin Z, Hu D, Chu R, Mo F, Liu C, Hiltunen E (2019) Effects of nitrogen source heterogeneity on nutrient removal and biodiesel production of mono- and mix-cultured microalgae Energ Convers. Manage 201:112144
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Author MPR express her gratitude to Mission innovation unit, Department of Biotechnology, New Delhi (INDIA) for financial support [file no.BT/PR31218/PBD/26/771/2019].
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Khanra, A., Vasistha, S., Kumar, P. et al. Role of C/N ratio on microalgae growth in mixotrophy and incorporation of titanium nanoparticles for cell flocculation and lipid enhancement in economical biodiesel application. 3 Biotech 10, 331 (2020). https://doi.org/10.1007/s13205-020-02323-0
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DOI: https://doi.org/10.1007/s13205-020-02323-0